GBM cells and glioma stem cells with EGFRvIII (+) significantly increased cell viability and cell migration. (A) The mRNA expression of EGFRvIII was detected by qRT-PCR in U87-EGFRvIII (+) cells, LN229-EGFRvIII (+) cells, and BNI-21-1-S-EGFRvIII (+) cells. (B) The protein expression of EGFRvIII was detected by Western blot. (C) The cell proliferation was detected by CCK-8 in U87-EGFRvIII (+) cells, LN229-EGFRvIII (+) cells, and BNI-21-1-S-EGFRvIII (+) cells. (D) The tumor growth of GBM in the orthotopic xenograft tumor model of mice was detected by in vivo bioluminescence imaging (n = 10 per group). (E) The cell migration was detected by Transwell in U87-EGFRvIII (+) cells, LN229-EGFRvIII (+) cells, and BNI-21-1-S-EGFRvIII (+) cells (scale bar: 100 μm). Representative data are determined from three independent experiments (n = 3 per group) vs. EGFRvIII (−). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 by Student’s two-tailed t-test.